Characterizing new redox roles for protein ubiquitination in human cells

表征人类细胞中蛋白质泛素化的新氧化还原作用

• 批准号: 10371928
• 负责人: Gustavo M Silva
• 金额: $ 42.15万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-05 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10371928
• 关键词: Affect; Aging; Antioxidants; Arsenicals; Atmosphere; Biological Assay; CRISPR screen; CRISPR/Cas technology; Cell Death; Cells; Cessation of life; Chronic; Chronic Disease; Chronic stress; Clustered Regularly Interspaced Short Palindromic Repeats; Degenerative Disorder; Deubiquitinating Enzyme; Diabetes Mellitus; Disease; Drug Targeting; Endoplasmic Reticulum; Enzymes; Exposure to; Foundations; Future; Gene Expression; Genes; Genomic Instability; Goals; Growth; Health; Human; Immunofluorescence Immunologic; Individual; Inflammation; Investigation; Lead; Learning; Libraries; Malignant Neoplasms; Mammalian Cell; Mediating; Metabolic; Methodology; Modification; Molecular; Monitor; Mutation; Nature; Nerve Degeneration; Oxidation-Reduction; Oxidative Stress; Pathway interactions; Pharmacology; Physiological; Polyribosomes; Process; Production; Protein Biosynthesis; Protein Dynamics; Protein Family; Proteins; Proteome; Proteomics; Regulation; Research; Resistance; Resolution; Ribosomes; Role; Signal Recognition Particle; Small Interfering RNA; Stress; System; Tertiary Protein Structure; Toxic Environmental Substances; Translations; Ubiquitin; Ubiquitination; Ultraviolet Rays; Water; Yeasts; acute stress; biological adaptation to stress; chemical genetics; design; drug development; endoplasmic reticulum stress; environmental stressor; genetic approach; genome editing; human disease; molecular imaging; novel; pollutant; prevent; protein aggregation; response; sodium arsenite; solar ultraviolet radiation; stressor; targeted treatment; trafficking; tumor progression

PROJECT SUMMARY Human cells are constantly subjected to oxidative stress due to the exposure to solar UV radiation and to pollutants in the water and in the atmosphere. To cope with the harms of oxidative stress that can lead to cellular malfunction and death, cells must reprogram gene expression and adjust the dynamics of protein production and degradation. Ubiquitin is a prominent protein modifier that controls the fate and the function of numerous proteins, and reshapes the proteome during environmental stresses. In response to oxidative stress, thousands of proteins accumulate ubiquitin modification and are involved in a variety of functions that support cellular survival. My lab discovered a new ubiquitin-dependent pathway that regulates protein production in yeast; however little is known about the importance and regulation of this pathway in human cells. In addition, to modify a large number of proteins with ubiquitin, human cells rely on hundreds of ubiquitin enzymes that selectively recognize their dedicated targets. However, the contribution of each ubiquitin enzyme to cellular resistance to stress is largely unknown. The purpose of the proposed project is to identify and characterize the roles of ubiquitin as a master regulator of the stress response. In Aim 1, we will investigate the molecular details of localized ubiquitin accumulation under oxidative stress, including when, why, and how it occurs. In Aim 2, we will investigate different aspects of cellular response to stress using genome editing methodologies targeting functional domains of > 700 ubiquitin enzymes. As oxidative stress is the underlying cause of a variety of human conditions such as aging, tumor progression, and degenerative disease, our research will provide a new understanding of ubiquitin function in the promotion of cellular health. Defining the importance of these pathways and the physiological role of ubiquitin enzymes will provide several avenues for drug development and it will allow us to learn the fundamental principles of the enzymatic regulation of protein ubiquitination during cellular resistance to stress.

项目概要 由于暴露在太阳紫外线辐射下，人体细胞不断遭受氧化应激 水和大气中的污染物。应对可能导致细胞损伤的氧化应激危害 功能障碍和死亡，细胞必须重新编程基因表达并调整蛋白质生产的动态和 降解。泛素是一种重要的蛋白质修饰剂，控制着多种蛋白质的命运和功能， 并在环境压力下重塑蛋白质组。为了应对氧化应激，数千种蛋白质 积累泛素修饰并参与支持细胞生存的多种功能。我的实验室 发现了一种新的泛素依赖性途径，可调节酵母中蛋白质的产生；然而我们知之甚少 关于该途径在人体细胞中的重要性和调节。另外，要修改大量 具有泛素的蛋白质，人类细胞依赖于数百种泛素酶来选择性地识别它们 专门的目标。然而，每种泛素酶对细胞抗应激能力的贡献在很大程度上是 未知。拟议项目的目的是确定和表征泛素作为主控的作用 应激反应的调节器。在目标 1 中，我们将研究局部泛素的分子细节 氧化应激下的积累，包括何时、为何以及如何发生。在目标 2 中，我们将调查 使用针对功能域的基因组编辑方法来研究细胞对应激反应的不同方面 > 700 种泛素酶。由于氧化应激是多种人类疾病的根本原因，例如 衰老、肿瘤进展和退行性疾病，我们的研究将为泛素提供新的认识 具有促进细胞健康的功能。定义这些途径的重要性和生理作用 泛素酶的研究将为药物开发提供多种途径，并使我们能够了解 细胞抵抗应激过程中蛋白质泛素化酶促调节的基本原理。